Leucocytic extract and method of making the same.



30:31am; A. AacnIBAnn, or oaxmmn, CALIFORNIA.

nnucocY'rIc nxraac'r AND METHOD or MAKING THE SAME.

No Drawing.

To all echo m. it may concern:

Be it known that I, Ronnn'r A. ARCHIBALD,

a citizen oft he United States, and a'resident of Oakland, county of .Alameda, and

5 State of California, have invented a certain .new and useful Leucocytic Extract and Method of Making the Same, of which the following is a specification.

The invention relates to leucocytic ex- 1 tract.

The object of the invention is a. provide a liquid or extract which when injected under the skin of a person having an infection or an infectious disorder or malady will produce a marked curative effect on the disorder or'malady or remove the infection.

Another object of the invention is to provide a method of making such extract. The invention possesses innumerable advantages inthe treatment and cure of infections, but I am .notsuiiicientlycertam as to the effects produced in the body by the inoculation of the extract tomake an absolute statement as to the biological or bac- 5- teriological changeswhich occur after the inoculation. I'am of .the opinion that the extract in some manner greatlyincreases the number of or the activity or capacity of the leucocytes or white corpuscles of the blood.

The leucocytes prey upon and take into their substance bacteria and other micro-organisms within the blood and tissues, and if their number or activity is increased, they operate to remove the infection. Whatever B the action of the extract is, I know from 'ex-' periment that it is extremely useful in the treatment of infections.

The extract is not necessarily limited inits production to the extract process herein 40 described, but may be produced by modifications of such process.

The extract is made from the blood of an animal such as the ox, hog, sheep, horse, dog or other domestic animal, the blood being in a normal condition,so that the leucocytes are normal and free from the influence of infection or inflammation. ,The blood is preferably obtained from the jugular vein of an animal by insertinga cannula into at-cleaned and 5 disinfectedarea, or may be..o'btained at a vent the blood-{from coagulating. When the blood arrives atfthe laboratory moreione slaughterhouse. The blood-from the animal is allowed to flow directlylinto sterile flasks which contain a suflicient amount of one per cent. sodium citrate solution to pre-' Specification of Iietterslatent.

Application filed July 18,

1914. Serial No. 850,591.

Patented Feb. 8, 1916.

per cent. sodium citrate solution is adde d in varying amounts to further dilute the. blood for the purpose of facilitating the precipitation of the blood corpuscles. The

amount ofsodium citrateadded at this time. depend upon the species of animal from ture is added a five-tenths of one per cent. acetic acld solution or other equivalent S0111- which the blood was obtained. "To this mix tion, for the purpose of breaking up and destroying the red blood corpuscles. The amount of the acetic acid solution added depends upon 'the character of blood'being treated. The 'mixture is then centrifuged in' a high powered centrifuge to precipitate the wh1te blood corpuscles. fluid which consists of the sodium citrate solution, the aceticacid solution and blood ,serum, is siphoned ofl and discarded. The

The supernatant Y sediment so obtained which is very rich in normal leucocytes is washedseveral times with physiological salt solution for the purpose of getting rid of the debris, acid and emoglobin, the result of the disintegration of the red blood corpuscles.

The washed leucocytic sediment is ground with quartz sand for the purpose of breaking up the masses of leucocytes and to assist in disintegrating the leucocytes themselves. This permits auto-digestion to progress with greater ease and rapidity and the solublesubstances contained within the cell walls of leucocytes are more easily extracted during the'process of auto-digestion. The acidity is then determined by titration,

using phenolphthalein, litmus or other material, as an indicator., Suflicient one tenth normal sodium hydrate solution is added to this emulsion to neutralize the acidity produced previously 'by the addition of .the

acetic acid solution. To this sediment distilled water is added in the proportion of about four volumes of water to one of sediment for thepurpose of dilution. The mixture is then exposed to a temperature. of

about 58 C. for approximately one hour. for

the purpose of destroying any antiferments' it may contain. It isthen placed in an incubator andmaintained at a temperature of about 37 (1,. until autodigestion, or lys 1s of the leucocytes is complete. .After the. leu- H cocytes have become broken up by reason of v this digestion, andthe protoplasm has been liberated, the mixture is centrifuged,-the' supernatent fluid or extract carefully .decanted, and 'suflicient trikresol or other preservative added-to it for preservative purposes. It is then sterilized by exposure to a temperature of 58 C. for one hour.

Each lot of extract is carefully examined 7 bacteriologically and biologically to deter mine if it is sterile'and its ability to produce certain definite blood changes is determined by the inoculation of definite quantibits and guinea pigs.

- ties into experimental, animals such as rab- Tli'e effect of the extract, when injected subcutaneously, is to increase thenumber of leucocytes and especially the polymophonuclear leucocytes which are believed, to be the most active of all of the white blood cells in the control of infectious diseases and other pathological conditions.

I claim:

1. Anextract obtained fromvtheliberated protoplasm of digested normal leucocytes in permanent solutionand possessing the'characteristic when injected parenterally of stimulating the production of a leucocytosis y and increasing t 1e activity of the individual leucocytes:

2. An extract in soluble form neutral, 1n

reaction which is obtainedifrom'the libe'rated protoplasm of digested normal leucocytes in permanent-solution and possessing the characteristic when injected parenterally of stimulating the production. of a leucocytosis and increasing the activity of the individual leucocytes.

3. The process of preparingleucocyt ic ex tract, which consists in separating the normal leucocytes from blood, diluting the resultant mass, incubating the same to facilitate a-utodigestion r lysis, and separating the liquid content from the mass.

-l. I he method of preparing leucocytic'ex tract, which consists in separating the'normal leucocytes from the blood, adding distilled water to said leucocytes, heating the mixture to destroy any antiferments it may contain, incubating the mixture to facilitate autodigestion or lysis 'of the leucocytes,

and then separating'the liquid content from the mass. b

5. The method of preparing l'eucocytic extract, which consists in adding a weak solution of acetic acid to blood, whereby the red blood corpusclesaredestroyed', centrifuging solution to blood for the purpose of destroying the red blood corpuscles, centrifuging the resultant mass to precipitate the leuco-' eytes, removing the supernatant fluid, washing the precipitate with salt solution,""""

emulsifying the washed precipitate, neutralizing its acidity wlth one-tenth normal sodium hydrate solution, adding distilled water to the mixture, exposingithe mixture to a temperature of about 58 centigrade for approximately one hour to destroy any antiferments it may contain, incubating the mixturefto facilitate autodigestion or lysis of the leucocytes, removing the supernatant fluid, sterilizing the fluid, and adding a preservative thereto.

In testimony whereof'I have hereunto set my hand at San Francisco, California, this 8th day of July 1914 ROBERT A.

i In presence of- H. G. Pnos'r, M. LE' CONTE.

ARCHIBALD. 

